Introduction to Organoids
Organoids are three-dimensional cell cultures grown in vitro that mimic the three-dimensional structure and physiological functions of normal (or diseased) organs or tissues in vivo through 3D cultivation. In simple terms, organoids are three-dimensional cell cultures where stem cells are cultured in matrix gel and, under the influence of chemical inhibitors/activators, cytokines, culture additives, and other substances, develop into tissue structures resembling corresponding organs.
Characteristics of Organoids
Organoids possess self-renewal capabilities and can maintain the physiological structure and function of the source tissue, earning them the title of "micro-organs in a dish." Leveraging the self-renewal, differentiation, and self-organization abilities of stem cells, organoids can be cryopreserved for use as biological repositories and can be infinitely expanded. Organoids are highly complex and, compared to 2D cell cultures, more closely resemble the in vivo state.
Figure 1 Organoid Culture of Human Colon Adenocarcinoma Cells
Applications of Organoids
Organoids offer several advantages in molecular and cellular biology analyses, bridging the gap between animal and cellular levels and providing better solutions for research in areas such as tumor studies, drug screening, and regenerative medicine. They have been widely applied in various research fields, including functional tissue induction, disease modeling, drug screening, anti-inflammatory testing, and clinical research, showing great potential in both basic research and translational applications.
With the continuous development of organoid culture systems and experimental techniques, organoids have been used to culture various tissues and organs, including intestines (small intestine/colon), stomach, liver, heart, lungs, prostate, pancreas, kidneys, mammary glands, brain-like structures, retinas, and inner ears.
Organoids derived from tumor stem cells have also begun to demonstrate tremendous potential in understanding the mechanisms of tumor occurrence and development, screening drug sensitivity, and promoting precision medicine and personalized diagnosis and treatment. Several studies published in Cell and Science have shown that organoids exhibit high sensitivity and specificity in predicting the effectiveness of anticancer drugs. Recently, tumor organoids have shown promise in predicting patient responses to cancer drugs and helping develop personalized treatment plans.
1. Research on Developmental Mechanisms: Organoids' differentiation capabilities can be utilized to study the embryonic development process and its mechanisms. The induction process regulated by signaling pathways such as Wnt and BMP can be used to study the development of organs such as brain-like structures, pancreas, and stomach.
2. Establishment of Disease Injury Models: Specific tissues or organs induced by organoids can be used to study specific disease models. The teams of Zhao Bing and Lin Xinhua have used human-derived organoid infection models to study the molecular mechanisms of SARS-CoV-2 infection and liver injury, providing important tools for research on the pathogenic mechanisms of SARS-CoV-2 and subsequent drug development.
The research group led by Deng Hongkui from Peking University School of Life Sciences used small molecules and cytokines to stimulate the construction of a novel intestinal organoid with injury regeneration characteristics—Hyper organoid. This organoid has the characteristics of long-term passaging and amplification while maintaining the genome, promoting colonic tissue repair, and alleviating pathological symptoms in acute colitis animal models.
3. Regenerative Medicine: Stem cell-derived organoids can repair or replace damaged or diseased tissues to restore normal tissue function, with extensive applications in cell therapy for various neurodegenerative diseases, diabetes, cardiovascular diseases, retinal diseases, and spinal cord injuries.
As a new treatment modality in the field of regenerative medicine—DA01, using small molecules SB-431542, LDN193189, CHIR-99021, and Y-27632, as well as Sonic Hedgehog (Shh) protein, to stimulate pluripotent stem cells to differentiate into dopaminergic neurons, and transplant them into the injured areas of the brains of late-stage Parkinson's disease patients, providing new directions and ideas for the treatment of this disease.
4. Drug Toxicity and Efficacy Testing: Organoids can be used to validate the pharmacotoxicity of new drugs in specific organs or tissues, providing data support for drug development.
Hyman kidney organoids can be used to validate the nephrotoxicity of Cisplatin.
5. Drug Screening: Organoids derived from stem cells can be used for in vitro testing of drug responses, providing theoretical support for drug screening.
Colon organoids can be used to study drug treatment strategies for CFTR mutation patients, and tumor organoids can be used to evaluate individualized drug treatments for patients.
The Developmental Process of Organoids
Sources of Organoids
Normal organoids mainly originate from stem cells, which include pluripotent stem cells (PSCs) and adult stem cells (ASCs). Pluripotent stem cells include embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). Compared to pluripotent stem cells, the advantage of adult stem cells is their simpler and faster modeling process, but the disadvantage is that the organoid structures they construct are relatively simple. Organoid structures constructed from pluripotent stem cells are more complex.
Organoid Culture
Summary of commonly used small molecules in organoid culture:very useful, don't forget to bookmark it!
1. Y-27632: A potent inhibitor of Rock, targeting p160ROCK and ROCK-II. Used in the initial culture medium for its role in enhancing cell viability and clone efficiency.
Recommended working concentration: 10 μM
2. SB-202190: An efficient inhibitor of p38 MAPK kinase, inducing differentiation of human embryonic stem cells into cardiomyocytes and promoting neural stem cell self-renewal.
Recommended dissolution concentration: 10 mg in 3.018 mL of DMSO to achieve 10 mM, stored at -20°C.
Recommended working concentration: 10 μM
3. CHIR-99021: A GSK-3 inhibitor, inducing differentiation of human embryonic stem cells into endoderm. Used in kidney and retina organoid cultures.
Recommended dissolution concentration: 5 mg in 3.58 mL of DMSO to achieve 3 mM, stored at -20°C.
Recommended working concentration: 3 μM
4. A 83-01: An inhibitor of the Activin/NODAL/TGF-β pathway, promoting self-renewal of cells in vitro. Used in liver, prostate, and breast organoid cultures.
Recommended dissolution concentration: 5 mg in 5.93 mL of DMSO to achieve 2 mM, stored at -20°C. (Note: This product is unstable in solution and should be prepared fresh.)
Recommended working concentration: 2 μM
5. Gastrin I: Used to prolong the survival time of organoids in intestinal and liver organoid cultures.
Recommended dissolution concentration: 1 mg in 2.38 mL of 1% ammonia solution to achieve 0.2 mM, stored at -20°C.
Recommended working concentration: 10 nM
6. Nicotinamide: A B3 vitamin involved in various enzymatic reactions, promoting differentiation of cells in organoid cultures.
Recommended dissolution concentration: 100 mg in 8.19 mL of H2O (or DMSO) to achieve 100 mM, stored at -20°C.
Recommended working concentration: 10 mM
7. Forskolin: Activates adenylate cyclase, elevating intracellular cAMP levels and inducing differentiation of various cell types.
Recommended working concentration: 1-10 μM
8. Prostaglandin E2: Regulates cell proliferation and differentiation, particularly in liver and prostate organoid cultures.
Recommended dissolution concentration: 1 mg in 0.28 mL of DMSO to achieve 10 mM, stored at -20°C.
Recommended working concentration: 500 nM
9. N-acetyl-L-Cysteine: An antioxidant precursor with anti-apoptotic properties, necessary for most organoid cultures.
Recommended dissolution concentration: 2 g in 24.51 mL of H2O (or DMSO) to achieve 500 mM, stored at -20°C.
Recommended working concentration: 1 mM
Related products
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Product Name |
Catalog Number |
Specifications |
|
Recombinant Human/Mouse/Rat Activin A Protein |
C230520 |
10 μg/100 μg/500 μg |
|
Recombinant Human/Mouse/Rat Activin A Protein |
C230308 |
10 μg/100 μg/500 μg |
|
Recombinant Human Dickkopf-related protein-1,His (Human DKK-1,His ) |
C230321 |
20 μg/100 μg/500 μg |
|
Recombinant Human EGF Protein，His Tag |
C230329 |
100 μg/500 μg |
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Recombinant Human bFGF/FGF-2 Protein |
C230295 |
10 μg/100 μg/500 μg |
|
Recombinant Human FGF-4 Protein |
C230346 |
5 μg/100 μg/500 μg |
|
Recombinant Human FGF-9 Protein, His tag |
C230347 |
5 μg/100 μg/500 μg |
|
Recombinant Human FGF-10 Protein, His Tag |
C230418 |
5 μg/100 μg/500 μg |
|
Recombinant Human GDNF Protein |
C230355 |
10 μg/100 μg/500 μg |
|
Recombinant Human Noggin Protein,His Tag |
C230462 |
5 μg/100 μg/500 μg |
|
Recombinant Human NRG1-beta1 Protein |
C230454 |
10 μg/100 μg/500 μg |
|
Recombinant Human R-Spondin-1 Protein,His (Human RSPO1,His ) |
C230484 |
100 μg/1 mg |
|
Recombinant Human Sonic Hedgehog N-Terminus (Human SHH) |
C230493 |
5 μg/100 μg/500 μg |
|
Recombinant Human BAFF Protein, His tag |
C230294 |
5 μg/100 μg/500 μg |
|
Recombinant Human sCD40 Ligand Protein, His tag |
C230487 |
10 μg/100 μg/500 μg |
|
Recombinant Human BMP-4 Protein |
C230310 |
10 μg/100 μg/500 μg |
|
Recombinant Human Hepatocyte Growth Factor (Human HGF) |
C230260 |
5 μg/100 μg/500 μg |
|
Recombinant Human BDNF Protein, Flag tag |
C230262 |
5 μg/20 μg/100 μg |
Mini Lecture: Small molecule compounds refer to compounds with a molecular weight less than 1000 Da (especially less than 500 Da) that exhibit biological activity. Unlike cytokines and proteins, small molecule compounds can penetrate cell membranes and exert corresponding biological functions within cells. Small molecule compounds have numerous target sites and have been widely used in many important research areas in life sciences, including stem cells, organoids, immunology, neurobiology, epigenetics, apoptosis, ion channels, oncology, and signal transduction.
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